Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
Not CRISPR data: the image illustrates antibody quality for visualizing CRISPR experiment results.
PDI Double Nickase Plasmid (h): sc-400676-NIC
- Target species: human
- 20 µg of transfection-ready, purified plasmid DNA; Suitable for up to 20 transfections
- PDI Double Nickase Plasmid (h) consists of a pair of plasmids each encoding a D10A mutated Cas9 nuclease and a target-specific 20 nt guide RNA (gRNA) designed to knockout gene expression with greater specificity than its CRISPR/Cas9 KO counterpart
- Paired gRNA sequences are offset by approximately 20 bp to allow for specific Cas9-mediated double nicking of the genomic DNA, which mimics a DSB
- One plasmid in the pair contains a puromycin-resistance gene for selection; the other plasmid in the pair contains a GFP marker to visually confirm transfection
- PDI Double Nickase Plasmid (h) and PDI Double Nickase Plasmid (h2) encode distinct paired gRNA designs targeting P4HB. One or both designs may be available
- Following transfection, gene knockout efficiency can be assayed by WB, IF or IHC using antibody: PDI Antibody (C-2): sc-74551
Ordering Information
| Product Name | Catalog # | UNIT | Price | Qty | FAVORITES | |
PDI Double Nickase Plasmid (h) | sc-400676-NIC | 20 µg | $410.00 | |||
PDI Double Nickase Plasmid (h2) | sc-400676-NIC-2 | 20 µg | $410.00 |
Human P4HB encodes protein disulfide isomerase (PDI), a multifunctional oxidoreductase and chaperone that catalyzes disulfide bond formation, reduction, and isomerization during protein folding in the endoplasmic reticulum. PDI supports ER proteostasis through the unfolded protein response and ER-associated degradation, and it participates in redox signaling via thiol–disulfide exchange reactions. Dysregulated P4HB/PDI activity has been linked to altered cellular stress responses, changes in secretory pathway capacity, and proteostasis imbalance observed across multiple disease-relevant contexts, including cancer biology and neurodegeneration models. As a core component of oxidative protein folding, PDI is frequently studied in pathways governing protein maturation, secretion, and redox homeostasis.
PDI Double Nickase Plasmid (h) consists of a matched pair of plasmids engineered for high-specificity editing of the P4HB locus in human cell lines. Each plasmid expresses a Cas9 D10A nickase and a distinct sgRNA targeting opposite DNA strands within P4HB. When directed to adjacent sites on opposite DNA strands, the two nickases generate offset single-strand nicks that together produce a staggered double-strand break, requiring coordinated on-target activity from both guides. The resulting DNA break is resolved by endogenous cellular repair pathways, most commonly through non-homologous end joining (NHEJ), leading to insertions or deletions that disrupt P4HB function. By requiring dual sgRNA engagement at the target locus, the double nicking approach enhances editing specificity and provides a complementary CRISPR strategy for applications where additional control over targeting precision is desired.
To support efficient identification of edited cells, one plasmid encodes GFP for fluorescent visualization of transfected populations, while the companion plasmid carries a puromycin resistance gene for antibiotic selection. Together, these features support efficient enrichment of co-transfected populations and simplify the validation of P4HB-disrupted clones.
For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.